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Snakes in the family Elapidae largely produce venoms rich in three-fnger toxins (3FTx) that bind to the α1 subunit of nicotinic acetylcholine receptors (nAChRs), impeding ion channel activity. These neurotoxins immobilize the prey by disrupting muscle contraction. Coral snakes of the genus Micrurus are specialist predators who produce many 3FTx, making them an interesting system for examining the coevolution of these toxins and their targets in prey animals. We used a bio-layer interferometry technique to measure the binding interaction between 15 Micrurus venoms and 12 taxon-specifc mimotopes designed to resemble the orthosteric binding region of the muscular nAChR subunit. We found that Micrurus venoms vary greatly in their potency on this assay and that this variation follows phylogenetic patterns rather than previously reported patterns of venom composition. The long-tailed Micrurus tend to have greater binding to nAChR orthosteric sites than their short-tailed relatives and we conclude this is the likely ancestral state. The repeated loss of this activity may be due to the evolution of 3FTx that bind to other regions of the nAChR. We also observed variations in the potency of the venoms depending on the taxon of the target mimotope. Rather than a pattern of prey-specifcity, we found that mimotopes modeled after snake nAChRs are less susceptible to Micrurus venoms and that this resistance is partly due to a characteristic tryptophan→serine mutation within the orthosteric site in all snake mimotopes. This resistance may be part of a Red Queen arms race between coral snakes and their prey.

RESUMO

Snakes in the family Elapidae largely produce venoms rich in three-fnger toxins (3FTx) that bind to the α1 subunit of nicotinic acetylcholine receptors (nAChRs), impeding ion channel activity. These neurotoxins immobilize the prey by disrupting muscle contraction. Coral snakes of the genus Micrurus are specialist predators who produce many 3FTx, making them an interesting system for examining the coevolution of these toxins and their targets in prey animals. We used a bio-layer interferometry technique to measure the binding interaction between 15 Micrurus venoms and 12 taxon-specifc mimotopes designed to resemble the orthosteric binding region of the muscular nAChR subunit. We found that Micrurus venoms vary greatly in their potency on this assay and that this variation follows phylogenetic patterns rather than previously reported patterns of venom composition. The long-tailed Micrurus tend to have greater binding to nAChR orthosteric sites than their short-tailed relatives and we conclude this is the likely ancestral state. The repeated loss of this activity may be due to the evolution of 3FTx that bind to other regions of the nAChR. We also observed variations in the potency of the venoms depending on the taxon of the target mimotope. Rather than a pattern of prey-specifcity, we found that mimotopes modeled after snake nAChRs are less susceptible to Micrurus venoms and that this resistance is partly due to a characteristic tryptophan→serine mutation within the orthosteric site in all snake mimotopes. This resistance may be part of a Red Queen arms race between coral snakes and their prey.

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BACKGROUND: B chromosomes are extra elements found in several eukaryote species. Usually, they do not express a phenotype in the host. However, advances in bioinformatics over the last decades have allowed us to describe several genes and molecular functions related to B chromosomes. These advances enable investigations of the relationship between the B chromosome and the host to understand how this element has been preserved in genomes. However, considering that transposable elements (TEs) are highly abundant in this supernumerary chromosome, there is a lack of knowledge concerning the dynamics of TE control in B-carrying cells. Thus, the present study characterized PIWI-interacting RNA (piRNA) clusters and pathways responsible for silencing the mobilization of TEs in gonads of the cichlid fish Astatotilapia latifasciata carrying the B chromosome. RESULTS: Through small RNA-seq and genome assembly, we predicted and annotated piRNA clusters in the A. latifasciata genome for the first time. We observed that these clusters had biased expression related to sex and the presence of the B chromosome. Furthermore, three piRNA clusters, named curupira, were identified in the B chromosome. Two of them were expressed exclusively in gonads of samples with the B chromosome. The composition of these curupira sequences was derived from LTR, LINE, and DNA elements, representing old and recent transposition events in the A. latifasciata genome and the B chromosome. The presence of the B chromosome also affected the expression of piRNA pathway genes. The mitochondrial cardiolipin hydrolase-like (pld6) gene is present in the B chromosome, as previously reported, and an increase in its expression was detected in gonads with the B chromosome. CONCLUSIONS: Due to the high abundance of TEs in the B chromosome, it was possible to investigate the origin of piRNA from these jumping genes. We hypothesize that the B chromosome has evolved its own genomic guardians to prevent uncontrolled TE mobilization. Furthermore, we also detected an expression bias in the presence of the B chromosome over A. latifasciata piRNA clusters and pathway genes.

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APAF1 is an autosomal recessive inherited mutation, associated with Holstein haplotype 1 (HH1) and characterized by a substitution of cytosine for a thymine (c.1741C>T) in chromosome 5. The mutation causes fetal and embryonic loss, between 60 and 200 days of gestation, and reduced conception rate. The ARMS-PCR is considered a simple and low-cost method to determine single nucleotide polymorphism (SNP) with no need for genetic sequencing of the animal genome. This study aimed to verify the allelic frequency of APAF1 mutation in Brazilian Holstein cattle. A total of 248 Holstein DNA samples (210 cows and 38 bulls) were analyzed, and synthetic genes were manufactured to validate the primers developed by the authors. All animals assessed in this study were classified as wild-type for APAF1 mutation. The primers and protocol developed for the ARMS-PCR technique work with 100% specificity and efficiency since the amplicon formations are as expected according to the genotypes. In conclusion, the mutation responsible for APAF1 was not detected in the Brazilian Holstein cattle population assessed in this prevalence study, although it is not possible to affirm that APAF1 does not occur in Brazilian Holstein animals. The tetra-primer ARMS-PCR protocol for APAF1 mutation that has been validated here may be a relatively simple and economical method to determine the animals' genotype.

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STAT2 plays a strategic role in defending viral infection through the signaling cascade involving the immune system initiated after type I interferon release. Many flaviviruses target the inactivation or degradation of STAT2 as a strategy to impair this host's line of defense. Primates are natural reservoirs for a range of disease-causing flaviviruses (e.g., Zika, Dengue, and Yellow Fever virus), while rodents appear less susceptible. We analyzed the STAT2 coding sequence of 28 Rodentia species and 49 Primates species. Original data from 19 Platyrrhini species were sequenced for the SH2 domain of STAT2 and included in the analysis. STAT2 has many sites whose variation can be explained by positive selection, measurement by two methods (PALM indicated 12, MEME 61). Both evolutionary tests significantly marked sites 127, 731, 739, 766, and 780. SH2 is under evolutionary constraint but presents episodic positive selection events within Rodentia: in one of them, a moderately radical change (serine > arginine) at position 638 is found in Peromyscus species, and can be implicated in the difference in susceptibility to flaviviruses within Rodentia. Some other positively selected sites are functional such as 5, 95, 203, 251, 782, and 829. Sites 251 and 287 regulate the signaling mediated by the JAK-STAT2 pathway, while 782 and 829 create a stable tertiary structure of STAT2, facilitating its connection with transcriptional co-activators. Only three positively selected sites, 5, 95, and 203, are recognized members who act on the interface between STAT2 and flaviviruses NS5 protein. We suggested that due to the higher evolutionary rate, rodents are, at this moment, taking some advantage in the battle against infections for some well-known Flaviviridae, in particular when compared to primates. Our results point to dynamics that fit with a molecular evolutionary scenario shaped by a thought-provoking virus-host arms race.

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Necator americanus is a worm that parasites the small intestine of humans and is highly prevalent in regions with poor sanitary conditions. The main strategy to control this helminth is by mass benzimidazole administration, however, periodic use of these drugs can select strains of parasites resistant to treatment. Single nucleotide polymorphisms (SNPs) in the beta-tubulin isotype 1 gene located at codons 167, 198 and 200 have been associated with benzimidazole resistance in some nematodes. Previously, our group detected the presence of some of these SNPs in populations of soil-transmitted helminths collected in different locations in Brazil. Here, we evaluated the SNP at codon 167, which has recently been shown to be associated with failure of benzimidazoles to treat N. americanus. Our ARMS-PCR analyses were performed using 524 single N. americanus eggs from 48 patients' feces collected in six Brazilian states; however, we did not detect any mutated samples at codon 167. This study builds on previous work, helping us monitor the presence of resistance-related genotypes in Brazilian helminth populations. The data presented here can assist in the implementation of future control strategies.

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Complex life cycle parasites can manipulate the behavior of intermediate hosts in order to reduce their fitness and increase the chance of completing life cycle. In order to understand the effects of the trematode parasites of the genus Clinostomum on host fish Loricariichthys platymetopon, a filmed experiment was carried out to quantify the foraging activity of hosts with different intensities of infection. The results suggest that hosts with higher parasite intensities reduced foraging activity early in the morning when compared to hosts with low intensities. This period may be critical for hosts since birds, the target hosts of such trematodes, forage intensively at dawn.

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In recent years, SE Brazil, the most populous region in the country with an estimated population of 88 million, has been experiencing an alarming increase in scorpions accidents (scorpionism), mainly caused by the yellow scorpion (Tityus serrulatus), or "escorpião amarelo" in Portuguese. This species is considered particularly dangerous to humans and can reproduce by parthenogenesis favouring rapid dispersal and colonization of new environments. Since the 1940s, owing to the growing danger represented by scorpionism, public control policies have been developed, including active search for scorpions, together with the use of toxic substances applied in places most likely to serve as their refuges. Even so, the number of accidents is increasing year by year, presently at an alarming rate. It seems evident that the increase in accidents is directly (or primarily) related to the lack of predators that in healthy environmental conditions would naturally control scorpion populations. However, due to environmental changes, leading to a lack of predators, scorpions have been gradually invading the urban environment. Arachnids and insects in general, as well as some other invertebrates, are preyed upon by anuran amphibians (toads, frogs and tree frogs). Toads (family Bufonidae) are nocturnal, large, and highly voracious animals, capable of actively exploring extensive areas and consuming large numbers of insects and arachnids daily. One of the most common toad species in southeastern Brazil is Rhinella icterica. Both R. icterica and T. serrulatus inhabit the same nocturnal environment. The predatory action of toads, specifically on scorpions, is practically unknown from behavioural and toxinological points of view. Thus, we studied the predatory behaviour of this toad against the yellow scorpion and evaluated the resistance of the amphibian to scorpion venom. Our results show that R. icterica is a voracious predator of T. serrulatus and is extremely resistant to its venom. Human/toad relationship throughout western history has always been very conflicted and possibly one of the factors that most has contributed to human ignorance of the role of these amphibians in maintaining ecological balance. Presently, the control of scorpionism is being performed through active search and/or the use of chemical agents, although showing little efficacy in reducing human accidents. In the medium or long term, more effective actions taking into account the biology of scorpions and their predators have never been taken to reduce these accidents.

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In recent years, SE Brazil, the most populous region in the country with an estimated population of 88 million, has been experiencing an alarming increase in scorpions accidents (scorpionism), mainly caused by the yellow scorpion (Tityus serrulatus), or "escorpião amarelo" in Portuguese. This species is considered particularly dangerous to humans and can reproduce by parthenogenesis favouring rapid dispersal and colonization of new environments. Since the 1940s, owing to the growing danger represented by scorpionism, public control policies have been developed, including active search for scorpions, together with the use of toxic substances applied in places most likely to serve as their refuges. Even so, the number of accidents is increasing year by year, presently at an alarming rate. It seems evident that the increase in accidents is directly (or primarily) related to the lack of predators that in healthy environmental conditions would naturally control scorpion populations. However, due to environmental changes, leading to a lack of predators, scorpions have been gradually invading the urban environment. Arachnids and insects in general, as well as some other invertebrates, are preyed upon by anuran amphibians (toads, frogs and tree frogs). Toads (family Bufonidae) are nocturnal, large, and highly voracious animals, capable of actively exploring extensive areas and consuming large numbers of insects and arachnids daily. One of the most common toad species in southeastern Brazil is Rhinella icterica. Both R. icterica and T. serrulatus inhabit the same nocturnal environment. The predatory action of toads, specifically on scorpions, is practically unknown from behavioural and toxinological points of view. Thus, we studied the predatory behaviour of this toad against the yellow scorpion and evaluated the resistance of the amphibian to scorpion venom. Our results show that R. icterica is a voracious predator of T. serrulatus and is extremely resistant to its venom. Human/toad relationship throughout western history has always been very conflicted and possibly one of the factors that most has contributed to human ignorance of the role of these amphibians in maintaining ecological balance. Presently, the control of scorpionism is being performed through active search and/or the use of chemical agents, although showing little efficacy in reducing human accidents. In the medium or long term, more effective actions taking into account the biology of scorpions and their predators have never been taken to reduce these accidents.

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Single nucleotide polymorphisms (SNPs) are the main type of variation in genome, enabling them to be associated with traits of economic importance in livestock. Genome-wide association studies (GWAS) have led to the discovery of SNPs associated with desirable traits in sheep. However, in these studies, SNPs are genotyped by high-throughput methods in genome scale, which are expensive and require sophisticated equipment and analysis methods. Therefore, the goal of this study was to develop a reliable, rapid, and inexpensive polymerase chain reaction (PCR)-based method to genotype a medium number of animals for a few candidate SNPs previously associated with desirable phenotypes in sheep by GWAS, using markers associated with gastrointestinal nematode resistance as a model. DNA extracted from white-blood cells of 150 sheep was submitted to PCR amplification followed by agarose gel electrophoresis and determination of banding pattern. Tetra-primer ARMS-PCR was successfully optimized after changes in annealing temperature; annealing and extension times; concentration of MgCl2 and DNA; ratios of inner, outer, forward and reverse primer; and addition of adjuvants, for genotyping the OAR2_14765360, OAR6_81718546, OAR11_62887032, and OAR12_69606944 SNPs in sheep. An extensive optimization of tetra-primer ARMS-PCR resulted in a suitable, simple, cost-effective PCR-based method of genotyping four SNP markers previously detected by chip arrays.

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The chemical control of Triatoma infestans, the major Chagas disease vector in southern South America, has been threatened in the last years by the emergence of pyrethroid-resistant bug populations. As an alternative approach, the efficacy of the entomopathogenic fungus Beauveria bassiana to control T. infestans populations (regardless their pyrethroid susceptibility) has been demonstrated. Growing research efforts on the interaction between T. infestans and B. bassiana by molecular, ecological, biochemical and behavioral traits has allowed framing such interaction as an evolutionary arms race. This review will focus on the relationships established in this particular host-pathogen system, compiling available data on the relevance of fungal pathogenesis, insect behavior, population dynamics and human intervention to favor fungal dissemination in bug populations. The current snapshot shows the fungus ahead in the evolutionary arms race and predicts a promissory landscape for the biological control of Chagas disease vectors.

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RESUMEN: Las variaciones anatómicas del músculo bíceps braquial en cuanto a la presencia de cabezas accesorias o supernumerarias pueden tener incidencia importante en los diagnósticos clínicos y radiológicos, así como en procedimientos quirúrgicos. La ausencia de una clasificación adecuada para las variaciones del músculo bíceps braquial y los hallazgos del presente estudio le permitieron a los autores presentar una propuesta de clasificación basada en el número de cabezas accesorias y en el origen de ellas. El objetivo del estudio fue determinar la frecuencia con que se presentan variaciones en el número de cabezas accesorias y proponer una clasificación para las variaciones del músculo bíceps braquial. En el estudio de tipo descriptivo, de corte transversal, se analizaron 90 brazos de 46 cadáveres colombianos. Se encontró una alta frecuencia (19,6 %) en las variaciones del músculo, 4,3 % bilateralmente y 15,2 % unilateralmente. En los 11 brazos (12,2 %) que presentaron la variación del músculo, la presencia de una sola cabeza accesoria fue la más alta (81,8 %), siendo el origen más frecuente en ellas el clasificado por los autores como 3b, 3 para la ubicación inferior medial y b para el intervalo entre la inserción del músculo coracobraquial y el origen del músculo braquial. Por lo detallado de la clasificación propuesta, se recomienda su uso cuando se reporten casos de variaciones anatómicas del músculo bíceps braquial. En los cadáveres de la población colombiana estudiados se encontró una alta frecuencia para la presencia de cabezas accesorias de este músculo, que amerita ser conocida por el personal médico.

SUMMARY: The anatomical variations of the biceps brachii muscle in relation to the number of supernumerary or accessory bellies may have important incidence in clinical and radiological diagnoses, as well as in surgical procedures. The absence of an appropriate classification for the variations that the biceps brachii muscle can present and the findings of the present study, allowed the authors to present a classification proposal based on the number of accessory bellies and their origin. The study aimed to determine with which frequency the variations in the number of accessory bellies of the biceps brachii muscle are present, and to propose a classification for these variations. In the crosssectional, descriptive study 90 arms of 46 Colombian cadavers were analyzed. A high frequency (19.6 %) in the variations of the muscle was found, 4.3 % bilaterally and 15.2 % unilaterally. In the 11 arms (12.2 %) that presented the variation, the presence of only one accessory belly was the highest frequency (81.8 %), being the most frequent origin for them the one classified by the authors as 3b, 3 for the inferomedial situation and b for the interval between the insertion of the coracobrachii muscle and the origin of the brachii muscle. Because the classification proposed is very detailed, its use is recommended whenever cases of biceps brachii muscle variations are reported. In the cadavers of the Colombian population studied, a high frequency was found for the presence of supernumerary or accessory heads of this muscle, which deserves to be known by medical professionals.

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Hosts counteract infections using two distinct defence strategies, resistance (reduction in pathogen fitness) and tolerance (limitation of infection damage). These strategies have been minimally investigated in multi-host systems, where they may vary across host species, entailing consequences both for hosts (virulence) and parasites (transmission). Comprehending the interplay among resistance, tolerance, virulence and parasite success is highly relevant for our understanding of the ecology and evolution of infectious and parasitic diseases. Our work investigated the interaction between an insect parasite and its most common bird host species, focusing on two relevant questions: (i) are defence strategies different between main and alternative hosts and, (ii) what are the consequences (virulence and parasite success) of different defence strategies? We conducted a matched field experiment and longitudinal studies at the host and the parasite levels under natural conditions, using a system comprising Philornis torquans flies and three bird hosts - the main host and two of the most frequently used alternative hosts. We found that main and alternative hosts have contrasting defence strategies, which gave rise in turn to contrasting virulence and parasite success. In the main bird host, minor loss of fitness, no detectable immune response, and high parasite success suggest a strategy of high tolerance and negligible resistance. Alternative hosts, on the contrary, resisted by mounting inflammatory responses, although with very different efficiency, which resulted in highly dissimilar parasite success and virulence. These results show clearly distinct defence strategies between main and alternative hosts in a natural multi-host system. They also highlight the importance of defence strategies in determining virulence and infection dynamics, and hint that defence efficiency is a crucial intervening element in these processes.

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The clinical effect of prednisone in metastatic castration-resistant prostate cancer (mCRPC) is unknown. We performed a pooled analysis of control arms of randomized controlled trials that had or had not administered single-agent prednisone. Randomized controlled trials with a control arm that included single-agent placebo (or no anticancer therapy) or single-agent prednisone (with or without placebo) were eligible for analysis. Patients receiving prednisone combined with other agents in the control arm were excluded. The trial characteristics, baseline demographic data, overall survival (OS), progression-free survival (PFS), prostate-specific antigen (PSA) response, Response Evaluation Criteria In Solid Tumors response, and toxicities were recorded. The effect of prednisone was investigated for significance in bivariate models, adjusting for age, pre- and post-docetaxel status, Eastern Cooperative Oncology Group performance status, and trial publication year. Eighteen trials were included; 9 had control arms that contained prednisone (n = 2831) and 9 did not (n = 2784). No significant differences were identified for OS or toxicities of any grade. A significantly greater PSA response rate (18.8% vs. 2.5%; P = .023) and a trend toward more frequent grade ≥ 3 fluid retention (1.0% vs. 0.4%; P = .097) was seen in the prednisone group. Prednisone was also significantly associated with PFS after adjusting for docetaxel status. Single-agent prednisone for mCRPC did not improve OS but was associated with a greater PSA response rate and PFS. Overall and grade ≥ 3 toxicities were not significantly different with prednisone. With the exception of concurrent use with abiraterone or for palliative purposes, the routine use of prednisone for mCRPC appears unnecessary.

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BACKGROUND: The entomopathogenic nematode Steinernema carpocapsae has been used worldwide as a biocontrol agent for insect pests, making it an interesting model for understanding parasite-host interactions. Two models propose that these interactions are co-evolutionary processes in such a way that equilibrium is never reached. In one model, known as "arms race", new alleles in relevant genes are fixed in both host and pathogens by directional positive selection, producing recurrent and alternating selective sweeps. In the other model, known as"trench warfare", persistent dynamic fluctuations in allele frequencies are sustained by balancing selection. There are some examples of genes evolving according to both models, however, it is not clear to what extent these interactions might alter genome-level evolutionary patterns and intraspecific diversity. Here we investigate some of these aspects by studying genomic variation in S. carpocapsae and other pathogenic and free-living nematodes from phylogenetic clades IV and V. RESULTS: To look for signatures of an arms-race dynamic, we conducted massive scans to detect directional positive selection in interspecific data. In free-living nematodes, we detected a significantly higher proportion of genes with sites under positive selection than in parasitic nematodes. However, in these genes, we found more enriched Gene Ontology terms in parasites. To detect possible effects of dynamic polymorphisms interactions we looked for signatures of balancing selection in intraspecific genomic data. The observed distribution of Tajima's D values in S. carpocapsae was more skewed to positive values and significantly different from the observed distribution in the free-living Caenorhabditis briggsae. Also, the proportion of significant positive values of Tajima's D was elevated in genes that were differentially expressed after induction with insect tissues as compared to both non-differentially expressed genes and the global scan. CONCLUSIONS: Our study provides a first portrait of the effects that lifestyle might have in shaping the patterns of selection at the genomic level. An arms-race between hosts and pathogens seems to be affecting specific genetic functions but not necessarily increasing the number of positively selected genes. Trench warfare dynamics seem to be acting more generally in the genome, likely focusing on genes responding to the interaction, rather than targeting specific genetic functions.

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The genus Echinococcus consists of parasites that have a life cycle with two mammalian hosts. Their larval stage, called the hydatid cyst, develops predominantly in the liver and lungs of intermediate hosts. The hydatid cyst is the causative agent of cystic hydatid disease and the species Echinococcus granulosus, G1 haplotype, is responsible for the vast majority of cases in humans, cattle and sheep. Protein characterization in hydatid cysts is essential for better understanding of the host-parasite relationship and the fertility process of Echinococcus. The aims of this work were the identification and quantitative comparison of proteins found in hydatid fluid from fertile and infertile cysts from E. granulosus, in order to highlight possible mechanisms involved in cyst fertility or infertility. Hydatid fluid samples containing proteins from both E. granulosus and Bos taurus were analysed by LC-MS/MS. Our proteomic analysis of fertile and infertile cysts allowed identification of a total of 498 proteins, of which 153 proteins were exclusively identified in the fertile cyst, 271 in the infertile cyst, and 74 in both. Functional in silico analysis allowed us to highlight some important aspects: (i) clues about the possible existence of an "arms race" involving parasite and host responses in fertile and infertile cysts; (ii) a number of proteins in hydatid fluid without functional annotation or with possible alternative functions; (iii) the presence of extracellular vesicles such as exosomes, which was confirmed by transmission electron microscopy.

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The tetraprimer ARMS-PCR technique is efficient for SNP detection and can be used to search for polymorphisms associated with drug resistance. However, the establishment of this methodology is not always straightforward because of the constraints on primer design due to the restrictions of the polymorphic regions. Here, we describe the standardization of the tetraprimer ARMS-PCR methodology for the detection of a SNP at codon 198 of the Ancylostoma caninum ß-tubulin gene. This SNP is associated with resistance to albendazole in various nematodes. The methodology was used to screen 327 individuals from 6 different locations. No mutation was found in any of the samples. This methodology will be useful for screening for the E198A SNP in the ß-tubulin gene of canine hookworms in a broader population to determine whether this SNP is associated with benzimidazole resistance in this species. The method could also be adapted for the analysis of other SNPs in other nematode species.

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